Journal: Oncology Letters

Article Title: Silymarin induces multiple myeloma cell apoptosis by inhibiting the JAK2/STAT3 signaling pathway

doi: 10.3892/ol.2025.15172

Figure Lengend Snippet: Silymarin inhibits the JAK2/STAT3 signaling pathway in multiple myeloma cells. mRNA expression levels of (A) Bcl-2 and (B) Bcl-xL in the RPMI 8226 cell line (n=3). mRNA expression levels of (C) Bcl-2 and (D) Bcl-xL in the H929 cell line (n=3). Immunofluorescence images of (E) JAK2 and (F) pJAK2. (G) Semi-quantification of levels of pJAK2 in the RPMI 8226 cell line (n=4). Immunofluorescence images of (H) STAT3 and (I) pSTAT3. Semi-quantification of (J) levels of pSTAT3 and (K) nuclear localization of pSTAT3 in the RPMI 8226 cell line (n=4). *P<0.05; **P<0.01; ***P<0.001. Error bars indicate standard deviation. p, phosphorylated; AU, arbitrary units.

Article Snippet: Subsequently, 5% bovine serum albumin (BSA; cat. no. A9418; MilliporeSigma) was applied to block nonspecific binding for 15 min. Primary antibodies targeting JAK2 (1:500; cat. no. PA5-11267; Thermo Fisher Scientific, Inc.), phosphorylated-JAK2 (1:500; cat. no. PA5-99341; Thermo Fisher Scientific, Inc.), STAT3 (1:100; cat. no. MA1-13042; Thermo Fisher Scientific, Inc.) and phosphorylated-STAT3 (1:100; cat. no. PA5-17876; Thermo Fisher Scientific, Inc.) were added to the cells and incubated overnight at 4°C.

Techniques: Expressing, Immunofluorescence, Standard Deviation

Journal: Oncology Letters

Article Title: Silymarin induces multiple myeloma cell apoptosis by inhibiting the JAK2/STAT3 signaling pathway

doi: 10.3892/ol.2025.15172

Figure Lengend Snippet: Interaction mode between silymarin and JAK2/STAT3. (A) Docking diagram of silymarin and JAK2 protein. (B) Docking diagram of silymarin and STAT3 protein. In addition, the planar structure of silymarin can be seen in the 2D image.

Article Snippet: Subsequently, 5% bovine serum albumin (BSA; cat. no. A9418; MilliporeSigma) was applied to block nonspecific binding for 15 min. Primary antibodies targeting JAK2 (1:500; cat. no. PA5-11267; Thermo Fisher Scientific, Inc.), phosphorylated-JAK2 (1:500; cat. no. PA5-99341; Thermo Fisher Scientific, Inc.), STAT3 (1:100; cat. no. MA1-13042; Thermo Fisher Scientific, Inc.) and phosphorylated-STAT3 (1:100; cat. no. PA5-17876; Thermo Fisher Scientific, Inc.) were added to the cells and incubated overnight at 4°C.

Techniques:

Journal: Investigative Ophthalmology & Visual Science

Article Title: Epac1 Inhibits Orbital Fibroblast Activation to Ameliorate Thyroid-Associated Orbitopathy-Like Features Through the JAK/STAT Signaling Pathway

doi: 10.1167/iovs.66.9.68

Figure Lengend Snippet: Epac1 affects the STAT3 signaling. ( A ) A schematic diagram of Epac1’s role in the JAK/STAT3 signaling pathway during the OF activation process. ( B ) TAO OFs transfected with sh- Epac1 or Epacl overexpression vector, treated with TGFβ1, and determined for the protein level of p-STAT3 and STAT3 using immunoblotting ( n = 3). One-way ANOVA with post hoc Tukey's test was used for analyzing comparisons among multiple groups. ** P < 0.01 versus NC-transfected OFs; ## P < 0.01 versus sh-NC-transfected OFs. ( C ) The protein level of p-STAT3 and STAT3 in TAO mouse OAT and OMT was determined using immunoblotting ( n = 3). One-way ANOVA with post hoc Tukey's test was used for analyzing comparisons among multiple groups. ** P < 0.01 versus healthy mice; # P < 0.05, ## P < 0.01 versus TAO + AAV-NC mice. TAO, thyroid-associated orbitopathy; OAT, orbital adipose tissues; OMT, orbital muscle tissues; AAV, adeno-associated virus; sh, short-hairpin RNA; NC, negative control.

Article Snippet: The used primary antibodies were α-SMA (55135-1-AP; Proteintech), Collagen I (14695-1-AP; Proteintech), Vimentin (10366-1-AP; Proteintech), Epac1 (DF6922; Affinity), fibronectin (15613-1-AP; Proteintech), collagen III (22734-1-AP; Proteintech), phosphorylated STAT3 (CSB-RA022812A727phHU; Cusabio, Wuhan, China), STAT3 (10253-2-AP; Proteintech), FABP4 (12802-1-AP; Protientech), and β-actin (CSB- MA000187 ; Cusabio).

Techniques: Activation Assay, Transfection, Over Expression, Plasmid Preparation, Western Blot, Virus, shRNA, Negative Control

Journal: Investigative Ophthalmology & Visual Science

Article Title: Epac1 Inhibits Orbital Fibroblast Activation to Ameliorate Thyroid-Associated Orbitopathy-Like Features Through the JAK/STAT Signaling Pathway

doi: 10.1167/iovs.66.9.68

Figure Lengend Snippet: STAT3 mediates the effects of Epac1 on TGFβ1-treated TAO OFs. ( A – G ) Under TGFβ1 treatment, TAO OFs were transfected with sh- Epac1 with or without the JAK-STAT pathway inhibitor Stattic (2 g/mL for 24 hours) and examined for cell viability using CCK-8 ( A ); cell migration using scratch wound healing and Transwell assays ( B , C ); α-SMA and fibronectin expressions using IF staining ( D ); the content of collagen I in cell supernatant using ELISA ( E ); the mRNA expression of Epac1 , α-SMA, vimentin, fibronectin, collagen I, and collagen III using qRT-PCR ( F ); the protein level of α-SMA, vimentin, fibronectin, collagen I, and collagen III using immunoblotting ( G ) ( n = 3). One-way ANOVA with post hoc Tukey's test was used for analyzing comparisons among multiple groups. ** P < 0.01 versus TGFβ1 + sh-NC; ## P < 0.01 versus TGFβ1 + sh- Epac1 + Stattic. sh, short-hairpin; NC, negative control; IF staining, immunofluorescent staining.

Article Snippet: The used primary antibodies were α-SMA (55135-1-AP; Proteintech), Collagen I (14695-1-AP; Proteintech), Vimentin (10366-1-AP; Proteintech), Epac1 (DF6922; Affinity), fibronectin (15613-1-AP; Proteintech), collagen III (22734-1-AP; Proteintech), phosphorylated STAT3 (CSB-RA022812A727phHU; Cusabio, Wuhan, China), STAT3 (10253-2-AP; Proteintech), FABP4 (12802-1-AP; Protientech), and β-actin (CSB- MA000187 ; Cusabio).

Techniques: Transfection, CCK-8 Assay, Migration, Staining, Enzyme-linked Immunosorbent Assay, Expressing, Quantitative RT-PCR, Western Blot, Negative Control